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Biomedical subjects

M Hans

Publications and source records attributed to M Hans.

At least 19 recordsLinked to original sources
PubMed1999

2-hydroxyglutaryl-CoA dehydratase from Clostridium symbiosum.

Component D (HgdAB) of 2-hydroxyglutaryl-CoA dehydratase from Clostridium symbiosum was purified to homogeneity. It is able to use component A from Acidaminococcus fermentans (HgdC) to initiate catalysis together with ATP, Mg2+ and a strong reducing agent such as Ti(III)citrate. Component D from C. symbiosum has a 6 x higher specific activity compared with that from A. fermentans and contains a second [4Fe-4S] cluster but the same amount of riboflavin 5'-phosphate (1.0 per heterodimeric enzyme, m = 100 kDa). Mössbauer spectroscopy revealed symmetric cube-type structures of the two [4Fe-4S]2+ clusters. EPR spectroscopy showed the resistance of the clusters to reducing agents, but detected a sharp signal at g = 2. 004 probably due to a stabilized flavin semiquinone. Three genes from C. symbiosum coding for components D (hgdA and hgdB) and A (hgdC) were cloned and sequenced. Primer extension experiments indicated that the genes are transcribed in the order hgdCAB from an operon only half the size of that from A. fermentans. Sequence comparisons detected a close relationship to the dehydratase system from A. fermentans and HgdA from Fusobacterium nucleatum, as well as to putative proteins of unknown function from Archaeoglobus fulgidus. Lower, but significant, identities were found with putative enzymes from several methanogenic Archaea and Escherichia coli, as well as with the mechanistically related benzoyl-CoA reductases from the Proteobacteria Rhodopseudomonas palustris and Thauera aromatica.

Amino Acid Sequence↗
PubMed1999

Functional consequences of mutations in the human alpha1A calcium channel subunit linked to familial hemiplegic migraine.

Mutations in alpha1A, the pore-forming subunit of P/Q-type calcium channels, are linked to several human diseases, including familial hemiplegic migraine (FHM). We introduced the four missense mutations linked to FHM into human alpha1A-2 subunits and investigated their functional consequences after expression in human embryonic kidney 293 cells. By combining single-channel and whole-cell patch-clamp recordings, we show that all four mutations affect both the biophysical properties and the density of functional channels. Mutation R192Q in the S4 segment of domain I increased the density of functional P/Q-type channels and their open probability. Mutation T666M in the pore loop of domain II decreased both the density of functional channels and their unitary conductance (from 20 to 11 pS). Mutations V714A and I1815L in the S6 segments of domains II and IV shifted the voltage range of activation toward more negative voltages, increased both the open probability and the rate of recovery from inactivation, and decreased the density of functional channels. Mutation V714A decreased the single-channel conductance to 16 pS. Strikingly, the reduction in single-channel conductance induced by mutations T666M and V714A was not observed in some patches or periods of activity, suggesting that the abnormal channel may switch on and off, perhaps depending on some unknown factor. Our data show that the FHM mutations can lead to both gain- and loss-of-function of human P/Q-type calcium channels.

Amino Acid Substitution↗
PubMed1999

Structural elements in domain IV that influence biophysical and pharmacological properties of human alpha1A-containing high-voltage-activated calcium channels.

We have cloned two splice variants of the human homolog of the alpha1A subunit of voltage-gated Ca2+ channels. The sequences of human alpha1A-1 and alpha1A-2 code for proteins of 2510 and 2662 amino acids, respectively. Human alpha1A-2alpha2bdeltabeta1b Ca2+ channels expressed in HEK293 cells activate rapidly (tau+10mV = 2.2 ms), deactivate rapidly (tau-90mV = 148 micros), inactivate slowly (tau+10mV = 690 ms), and have peak currents at a potential of +10 mV with 15 mM Ba2+ as charge carrier. In HEK293 cells transient expression of Ca2+ channels containing alpha1A/B(f), an alpha1A subunit containing a 112 amino acid segment of alpha1B-1 sequence in the IVS3-IVSS1 region, resulted in Ba2+ currents that were 30-fold larger compared to wild-type (wt) alpha1A-2-containing Ca2+ channels, and had inactivation kinetics similar to those of alpha1B-1-containing Ca2+ channels. Cells transiently transfected with alpha1A/B(f)alpha2bdeltabeta1b expressed higher levels of the alpha1, alpha2bdelta, and beta1b subunit polypeptides as detected by immunoblot analysis. By mutation analysis we identified two locations in domain IV within the extracellular loops S3-S4 (N1655P1656) and S5-SS1 (E1740) that influence the biophysical properties of alpha1A. alpha1AE1740R resulted in a threefold increase in current magnitude, a -10 mV shift in steady-state inactivation, and an altered Ba2+ current inactivation, but did not affect ion selectivity. The deletion mutant alpha1ADeltaNP shifted steady-state inactivation by -20 mV and increased the fast component of current inactivation twofold. The potency and rate of block by omega-Aga IVA was increased with alpha1ADeltaNP. These results demonstrate that the IVS3-S4 and IVS5-SS1 linkers play an essential role in determining multiple biophysical and pharmacological properties of alpha1A-containing Ca2+ channels.

Amino Acid Sequence↗
PubMed1999

Structure and functional characterization of a novel human low-voltage activated calcium channel.

We have isolated and characterized overlapping cDNAs encoding a novel, voltage-gated Ca2+ channel alpha1 subunit, alpha1H, from a human medullary thyroid carcinoma cell line. The alpha1H subunit is structurally similar to previously described alpha1 subunits. Northern blot analysis indicates that alpha1H mRNA is expressed throughout the brain, primarily in the amygdala, caudate nucleus, and putamen, as well as in several nonneuronal tissues, with relatively high levels in the liver, kidney, and heart. Ba2+ currents recorded from human embryonic kidney 293 cells transiently expressing alpha1H activated at relatively hyperpolarized potentials (-50 mV), rapidly inactivated (tau = 17 ms), and slowly deactivated. Similar results were observed in Xenopus oocytes expressing alpha1H. Single-channel measurements in human embryonic kidney 293 cells revealed a single-channel conductance of approximately 9 pS. These channels are blocked by Ni2+ (IC50 = 6.6 microM) and the T-type channel antagonists mibefradil (approximately 50% block at 1 microM) and amiloride (IC50 = 167 microM). Thus, alpha1H-containing channels exhibit biophysical and pharmacological properties characteristic of low voltage-activated, or T-type, Ca2+ channels.

3-Pyridinecarboxylic acid, 1,4-dihydro-2,6-dimethy↗
PubMed1998

Average African American three-dimensional computed tomography skull images: the potential clinical importance of ethnicity and sex.

The production of average 'normative' three-dimensional (3D) computed tomography surface images of the bony skull has only recently been explored. The authors wish to determine the effect of using sex- and ethnicity-specific adult average 3D skull images for comparisons with patient images at various stages of craniofacial surgical management (i.e., diagnosis, treatment planning, prosthetic design, image-guided operative procedures, and outcomes assessment). Craniofacial surgical reconstruction for abnormal patterns of development, cancer resection, or trauma are most likely to benefit from these comparisons. To morphometrically test the significance of separating normative 3D skull data by sex and ethnicity, the authors collected 52 3D, anatomical landmarks from 3D computed tomography scans of dry skulls of 20 Americans of European ethnicity and 20 Americans of primarily African (i.e., primarily African and some European) ethnicity. A Procrustes-based morphometric analysis of shape detects 1.2 times as much interethnic variance as intersex variance. The African American sample presents 4.2% more dolichocephaly, wider orbits, flatter nasal area, larger gnathic anatomy, and more procumbent dentition. Pooling the sexes across both groups, it is seen that men tend to have less bulbous crania, more protruding brows, noses, and masticatory muscle attachments, and relatively less protrusive palettes and anterior mandibles. Despite a small sample size (N = 40), the authors' results are statistically significant (P approximately 0.001 overall) for both of the main factors, sex and ethnicity, separately.

Adolescent↗
PubMed1997

Contribution of craniofacial risk factors in increasing apneic activity among obese and nonobese habitual snorers.

STUDY OBJECTIVE: To determine the role of craniofacial risk factors in increasing apneic activity between nonobese and obese habitual snorers. DESIGN: Cross-sectional. SETTING: Care-seeking volunteers identified through advertisements, and referral-based volunteers from two sleep centers serving the greater Cleveland area. PATIENTS: The study included 142 habitual snorers (mean +/- SD, 45.5 +/- 10.6 years, 68% men; 20% African-Americans) MEASUREMENTS AND RESULTS: Apneic activity was determined using unattended home sleep monitoring to assess the respiratory disturbance index (RDI). Independent variables included body mass index, cranial index, facial index, and 13 anatomic variables (lateral and frontal cephalometric radiographs were used to characterize craniofacial hard and soft tissues). A linear regression model explained approximately 54% and 53% of the variation in RDI (log transformed) scores for the nonobese and obese groups, respectively. The largest predictor of RDI in the nonobese group was tongue length, followed by alignment of the middle cranial fossa, and age. In the obese group, the largest predictor of RDI was hyoid to mandibular plane, followed by tongue length. CONCLUSIONS: Measurements of soft tissues of the oropharynx (especially the tongue) are more closely associated with increased apneic activity. In addition, the hard tissue anatomic limits of the oropharynx may place nonobese individuals in the at-risk group. Therefore, anatomic relationships that are temporally stable may be useful to predict apneic activity in later years.

Adult↗
PubMed1995

Activation of Ca2+ signaling in neutrophils by the mast cell-released immunophilin FKBP12.

The immunophilins of the FK506-binding protein (FKBP) family are intracellular proteins that bind the immunosuppresants FK506 and rapamycin. In this study we show that HMC-1 mast cells sensitized with IgE release FKBP12 upon stimulation with anti-IgE. The release is rapid and not affected by actinomycin D or cycloheximide, suggesting that it is due to exocytosis from a storage compartment. FKBP12 from HMC-1 mast cells exhibits biological activity. When applied extracellularly to human neutrophils, it induces transient changes in the intracellular Ca2+ concentration ([Ca2+]i) due to Ca2+ release from intracellular stores. Inhibition of [Ca2+]i changes by ruthenium red and ryanodine indicates that ryanodine receptor/Ca2+ release channels are involved in FKBP12-induced Ca2+ signaling. Neutrophil activation by mast cell-derived FKBP12 is prevented by complexing FKBP12 with FK506 or rapamycin. These results demonstrate that extracellular FKBP12 functions as a cytokine in cell-to-cell communication. They further suggest a pathophysiological role for FKBP12 as a mediator in immediate or type I hypersensitivity and may have implications for novel therapeutic strategies in the treatment of allergic disorders with FK506 and rapamycin.

Amino Acid Sequence↗
PubMed1995

Characteristics of a human N-type calcium channel expressed in HEK293 cells.

The human alpha 1B-1 alpha 2b beta 1-2 Ca2+ channel was stably expressed in HEK293 cells producing a human brain N-type voltage-dependent calcium channel (VDCC). Whole cell voltage-clamp electrophysiology and fura-2 based microfluorimetry have been used to study its characteristics. Calcium currents (ICa) recorded in transfected HEK293 cells were activated at potentials more depolarized than -20 mV with peak currents occurring at approx + 10 mV in 5 mM extracellular CaCl2. ICa and associated rises in intracellular free calcium concentrations ([Ca2+]i) were sensitive to changes in both the [Ca2+]o and holding potential. Steady-state inactivation was half maximal at a holding potential of -60 mV. Ba2+ was a more effective charge carrier than Ca2+ through the alpha 1B-1 alpha 2b beta 1-2 Ca2+ channel and combinations of both Ba2+ and Ca2+ as charge carriers resulted in the anomalous mole fraction effect. Ca2+ influx into transfected HEK293 cells was irreversibly inhibited by omega-conotoxin-GVIA (omega-CgTx-GVIA; 10 nM-1 microM) and omega-conotoxin-MVIIA; 100 nM-1 microM) whereas 1 microM) whereas no reductions were seen with agents which block P or L-type Ca2+ channels. The inorganic ions, gadolinium (Gd3+), cadmium (Cd2+) and nickel (Ni2+) reduced the ICa under voltage-clamp conditions in a concentration-dependent manner. The order of potency of the three ions was Gd3+ > Cd2+ > Ni2+. These experiments suggest that the cloned and expressed alpha 1B-1 alpha 2b beta 1-2 Ca2+ channel subunits form channels in HEK293 cells that exhibit properties consistent with the activity of the native-N-type VDCC previously described in neurons.

Cadmium↗
PubMed1994

Structure and functional characterization of neuronal alpha 1E calcium channel subtypes.

We have cloned overlapping cDNAs encoding alpha 1E Ca2+ channel subunits from mouse and human brain. We observed that these alpha 1E transcripts were widely distributed in the central nervous system. We also demonstrated the existence of two variants of the human alpha 1E subunit. Comparison of the sequence of these alpha 1E subunits to those from other species suggests that at least four alternatively spliced variants of alpha 1E exist. Expression of human alpha 1E in HEK293 cells and Xenopus oocytes produced high voltage-activated Ca2+ currents that inactivated rapidly (tau approximately 20 ms at 0 mV). The size of the currents obtained were enhanced approximately 40-fold by co-expression with human neuronal alpha 2 and beta Ca2+ channel subunits. alpha 1E currents were insensitive to the drugs and toxins previously used to define other classes of voltage-activated Ca2+ channels. Thus, alpha 1E-mediated Ca2+ channels appear to be a pharmacologically distinct class of voltage-activated Ca2+ channels.

Alternative Splicing↗
PubMed1994

Molecular pathways for presynaptic calcium signaling.

The results presented in this article describe two distinct, Ca-regulated molecular pathways in presynaptic terminals and implicate these two pathways in differentially mediating neurotransmitter secretion and PTP. Our current view of the Ca-dependent triggering of secretion and PTP is shown in Fig. 9. According to this scheme, differential activation of these two pathways is achieved by a combination of diffusion-based dilution of Ca that enters the terminal through voltage-gated Ca channels and by coupling these pathways to Ca receptors with different affinities for Ca ions. A simple way to achieve these conditions is to position these two receptors at different distances from the Ca channels, as shown in Fig. 2. Given that Ca ions are involved in activating many different presynaptic processes (Fig. 1), we propose that closer scrutiny of the molecular physiology of nerve terminals will reveal a wide variety of Ca-activated pathways responsible for producing these diverse processes.

Amino Acid Sequence↗
PubMed1992

The effects of the Le Fort I osteotomy on the periodontium.

Two age-matched populations of equal size (n = 40), one having orthodontic therapy and the other combined orthodontic therapy and orthognathic surgery, were evaluated for their periodontal status 1 to 10 years posttherapy. The parameters investigated were plaque index, gingival index, tooth mobility, width of keratinized tissue, probing depth, gingival recession, and attachment level. No significant differences were found (P less than .05). Within the surgery group, patients with maxillary osteotomies segmentalized between the central incisors (n = 11) and between the canines and second premolars (n = 12) were evaluated using the same parameters and compared with their nonsegmental counterparts. No significant differences were found for patients with osteotomies segmentalized between the central incisors. However, a statistically significant increase in probe depth and loss of attachment level of up to 0.3 mm was found at the sites of osteotomies segmentalized between the canine and second premolar (P less than .05). This difference was not considered clinically significant.

Adolescent↗
PubMed1992

Presynaptic calcium signals during neurotransmitter release: detection with fluorescent indicators and other calcium chelators.

Synthetic calcium buffers, including fluorescent calcium indicators, were microinjected into squid 'giant' presynaptic nerve terminals to investigate the calcium signal that triggers neurotransmitter secretion. Digital imaging methods, applied in conjunction with the fluorescent calcium indicator dye fura-2, reveal that transient rises in presynaptic calcium concentration are associated with action potentials. Transmitter release terminates within 1-2 ms after a train of action potentials, even though presynaptic calcium concentration remains at micromolar levels for many seconds longer. Microinjection of the calcium buffer, EGTA, into the presynaptic terminal has no effect on transmitter release evoked by single presynaptic action potentials. EGTA injection does, however, block the change in calcium concentration measured by fura-2. Therefore, the calcium signal measured by fura-2 is not responsible for triggering release. These results suggest that the rise in presynaptic calcium concentration that triggers release must be highly localized to escape detection with fura-2 imaging. Unlike EGTA, microinjection of BAPTA--a calcium buffer with an equilibrium affinity for calcium similar to that of EGTA--produces a potent, dose-dependent, and reversible block of action-potential evoked transmitter release. The superior ability of BAPTA to block transmitter release apparently is due to the more rapid calcium-binding kinetics of BAPTA compared to EGTA. Because EGTA should bind calcium within a few tens of microseconds under the conditions of our experiments, the inability of EGTA to block release indicates that transmitter release is triggered within a few tens of microseconds after the entry of calcium into the presynaptic terminal.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1991

Role of residual calcium in synaptic depression and posttetanic potentiation: fast and slow calcium signaling in nerve terminals.

Trains of action potentials evoked rises in presynaptic Ca2+ concentration ([Ca2+]i) at the squid giant synapse. These increases in [Ca2+]i were spatially nonuniform during the trains, but rapidly equilibrated after the trains and slowly declined over hundreds of seconds. The trains also elicited synaptic depression and augmentation, both of which developed during stimulation and declined within a few seconds afterward. Microinjection of the Ca2+ buffer EGTA into presynaptic terminals had no effect on transmitter release or synaptic depression. However, EGTA injection effectively blocked both the persistent Ca2+ signals and augmentation. These results suggest that transmitter release is triggered by a large, brief, and sharply localized rise in [Ca2+]i, while augmentation is produced by a smaller, slower, and more diffuse rise in [Ca2+]i.

Action Potentials↗
PubMed1990

The blocking effects of omega-conotoxin on Ca current in bovine chromaffin cells.

The effects of omega-conotoxin (omega-CgTX) on voltage-sensitive Ca currents (ICa) were studied in cultured bovine adrenal chromaffin cells. A maximal block of ICa of 40-50% was obtained with omega-CgTX in the microM range, and was independent of the holding potential. The onset of block was both concentration- and time-dependent. In bovine chromaffin cells, Ca channels, both sensitive and insensitive to omega-CgTX, appear to be present.

Adrenal Medulla↗
PubMed1990

Nicotinic cholinergic modulation of voltage-dependent calcium current in bovine adrenal chromaffin cells.

1. The effects of cholinergic agonists on voltage-dependent calcium current (ICa) were studied in cultured chromaffin cells from bovine adrenal medulla. 2. Application of both acetylcholine (ACh) and nicotine resulted in inward nicotinic current from a holding potential of -90 mV, and at the same time reversible decreases in depolarization-activated ICa. Both of these effects were blocked by d-tubocurarine, while atropine pre-treatment was ineffective. 3. Internal accumulation of neither Na+ nor Ca2+ seems likely to explain the nicotinic-agonist-dependent decrease in ICa, as the modulation was observed with symmetrical Na+ solutions, with Ca2(+)-free Ba2(+)-containing external solutions, from holding potentials of both -90 and -40 mV, and when the internal Ca2+ buffer capacity was increased. 4. Isodihydrohistrionicotoxin, an open-channel blocker which does not compete for the agonist binding site, completely inhibited inward cholinergic currents while the agonist-dependent decrease in ICa was seen in only two of fifteen cells. 5. The nicotinic agonist-mediated decreases in ICa were not voltage-dependent. 6. No changes in voltage-dependent INa were seen with the nicotinic agonists. 7. Muscarine, with or without GTP in the pipette solution, produced neither modulation of ICa nor any changes in steady holding currents. The nicotinic current and the reversible decrease in ICa induced by ACh and nicotine were not affected by including GTP, or the guanine nucleotide analogues GDP-beta-S and GTP-gamma-S, in the pipette solution. 8. A 10 min pre-incubation of the cells in a high-K+ solution optimal for catecholamine secretion did not affect the nicotinic agonist-mediated decreases in ICa.

Adrenal Medulla↗
PubMed1978

Infinite dilution conductimetry of plasma and urine: correlation with osmolality.

The infinite dilution conductivity (IDC) of plasma and urine allows a measurement of the electrolyte content in small samples (5 to 15 microliter). The method was compared to the corrected osmolality (II'p) measured by the freezing-point depression. A linear correlation existed between II'p and the IDC: for plasma: II'p = 13.10 sigma o,p + 37.00 (n = 46 and r = 0.9949) for urine: II'u = 12.75 sigma o,u + 16.56 (n = 85 and r = 0.9504). The measurement of the IDC does not depend on protein concentration and can be used instead of the osmometer methods to determine the total plasma and urine electrolyte content.

Blood Physiological Phenomena↗